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Biomedical subjects

D W Walker

Publications and source records attributed to D W Walker.

At least 37 records · Page 2Linked to original sources
PubMed1999

The development and pharmacological characterization of calcium channel currents in cultured embryonic rat septal cells.

We characterized the development and pharmacology of Ca(2+) channel currents in NGF-treated embryonic day 21 cultured rat septal cells. Using standard whole-cell voltage clamp techniques, cells were held at -80 mV and depolarized to construct current-voltage relations in conditions that eliminated Na(+) or K(+) currents. Barium (10 mM) was used as the charge carrier. Maximum current was produced when cells were depolarized to 0 or +10 mV. Recordings from 77 cells revealed that Ca(2+) channel current density increases over time in culture from nearly 0 pA/pF on day 2 in vitro (0.65+/-0.65 pA/pF) to (6.95+/-1.59 pA/pF) on days 6-8. This was followed by a period where currents became nearly 3 times more dense (21.05+/-7.16 pA/pF) at days 9-17. There was little or no evidence for low voltage activated currents. Bath application of 50-100 microM CdCl(2) abolished approximately 95% of the current. Application of 10 microM nimodipine produced a 50.5+/-3.22% reduction in current, 2 microM omega-CTx-GVIA produced a 32.4+/-7.3% reduction, and application of 4 microM omega-Aga-IVA produced a 29.5+/-5.73% reduction in current. When all three inhibitors (10 microM nimodipine, 2 microM omega-CTx-GVIA, and 4 microM omega-Aga-IVA) were applied simultaneously, a residual current remained that was 18.0+/-4.9% of the total current and was completely abolished by application of CdCl(2). This is the first report to characterize Ca(2+) channel currents in cultured embryonic septal cells. These data indicate that there is a steady increase in Ca(2+) channel expression over time in vitro, and show that like other cultured neuronal cells, septal cells express multiple Ca(2+) channel types including L, N, P/Q and R-type channels.

Animals↗
PubMed1999

Characterization of pontine neurons which respond to hypoxia in fetal sheep.

Hypoxia causes apnea and postural muscle hypotonia in fetal sheep, a response thought to arise by descending inhibition from a group of lateral pontine neurons that express FOS protein after hypoxia. To determine the neurochemical phenotype, and whether these neurons project to the cervical spinal cord, the retrograde tracer CTB-gold was injected into the C5-C8 ventral horn of four fetal sheep at 110 days gestation. Then, at 135 days each fetus was made hypoxic for 2 h by allowing the mother to breathe 7-8% O2. Immunocytochemistry showed that FOS-positive neurons in the subcoeruleus and Kolliker-Fuse regions of the pons were catecholaminergic, but not cholinergic or GABAergic, and a proportion of them contained CTB-gold particles, indicating direct connection with the cervical spinal cord. We suggest that these pontine neurons inhibit respiratory and postural muscle activities during hypoxia in fetal sheep.

Animals↗
PubMed1999

The distribution of FOS-immunoreactive neurons in the brainstem, midbrain and diencephalon of fetal sheep in response to acute hypoxia in mid and late gestation.

FOS immunohistochemistry was used to map the distribution of neuronal pathways activated by hypoxia in fetal sheep. Conscious pregnant sheep were exposed to hypoxia (7-9% O2, 1-2% CO2, balance N2) for 2 h at either 100-105 days (n=5) or 130-133 days gestation (n=5); term is approximately 147 days. The hypoxia caused cessation of breathing movements at both fetal ages, and increased FOS staining in the medulla (area postrema, dorsal motor nucleus of vagus, nucleus solitary tract, ventrolateral medulla); pons (locus coeruleus and subcoeruleus, lateral and medial parabrachial nuclei); midbrain (habenula, periaqueductal grey, substantia nigra, areas ventrolateral to Red Nucleus); and hypothalamus (anterior and lateral hypothalamic areas, paraventricular and supraoptic nuclei). The results were essentially the same at both gestational ages, except that hypoxia increased FOS-staining in the habenula only in the older fetuses. The presence of FOS protein in pontomedullary cardiorespiratory nuclei at 100-105 days gestation indicates that the peripheral chemoreceptors respond to hypoxia at this early age, and in the subcoeruleus and medial parabrachial regions of the pons is consistent with lesion studies suggesting these areas mediate the inhibition of fetal breathing in response to hypoxia. FOS staining in the ventrolateral periaqueductal grey and habenula was unexpected, and suggests that pathways normally involved in response to noxious stimuli, or which are part of the hypothalamic 'defense' response are activated by hypoxia in the fetus. Some FOS-labelling could arise secondarily as a consequence of the cardiovascular and endocrine responses to hypoxia.

Animals↗
PubMed1999

Tryptophan metabolism in pregnant sheep: increased fetal kynurenine production in response to maternal tryptophan loading.

OBJECTIVE: The effects of a tryptophan load on the plasma concentration of kynurenine, the precursor for the production in the brain of the neuroactive products kynurenic acid and quinolinic acid, were determined in pregnant sheep at midgestation and late gestation and in nonpregnant sheep. STUDY DESIGN: Pregnant ewes were given an intravenous infusion of 100 mg/kg L-tryptophan during 2 hours at 95 to 98 days' gestation (n = 4) or 135 to 138 days' gestation (n = 10). Nonpregnant ewes (n = 6) were studied in late estrus. Arterial blood samples taken from 2 hours before to 48 hours after the start of the infusion were used for analysis of plasma tryptophan, kynurenine, and cortisol concentrations. RESULTS: Tryptophan loading at both gestational ages resulted in significantly greater increases in kynurenine concentrations in fetal plasma (at 95-98 days' gestation, from 5.7 +/- 1.2 micromol/L [baseline] to 247.9 +/- 86.7 micromol/L (peak); at 135-138 days' gestation, from 9.0 +/- 2.3 micromol/L [baseline] to 289.0 +/- 194.0 micromol/L [peak]) than in maternal plasma [at 95-98 days' gestation, from 4.6 +/- 0.8 micromol/L [baseline] to 118.0 +/- 79.7 micromol/L [peak]; at 135-138 days' gestation, from 4.8 +/- 2.9 micromol/L [baseline] to 98.3 +/- 67.8 micromol/L [peak]). It took longer for kynurenine concentrations to return to basal values in the fetus (24-30 hours) than in the ewe (8-12 hours). The kynurenine responses in pregnant and nonpregnant ewes were not different from each other. CONCLUSION: The production of kynurenine from tryptophan is significantly greater in the fetal lamb than in the pregnant or nonpregnant adult ewe.

Animals↗
PubMed1999

Effect of nitric oxide synthase inhibition on the uterine vasculature of the late-pregnant ewe.

OBJECTIVES: We studied the role of nitric oxide in the maintenance of uterine vascular tone during pregnancy. STUDY DESIGN: Late-pregnant ewes were instrumented with ultrasonographic flow probes on the left and right main uterine arteries. A catheter was passed retrogradely into 1 uterine artery from a tributary. In 14 animals nitric oxide synthase inhibitor (N omega-nitro-L-arginine methyl ester; L-NAME) was infused at 2 or 20 mg/kg during a 1-minute period into either the main left or right uterine artery. RESULTS: The nitric oxide synthase inhibitor (20 mg/kg), infused into 1 uterine artery, significantly decreased uterine blood flow (P <.001) bilaterally, increased (P <.05) mean arterial pressure, and decreased heart rate (P <.05). The compound (when infused at 2 mg/kg) also significantly (P <.05) decreased uterine blood flow in the artery ipsilateral to, but not contralateral to, the infusion, with no change in mean arterial pressure and a decrease (P <.05) in heart rate. CONCLUSION: During ovine pregnancy, endogenous nitric oxide production contributes to uterine vasodilatation.

Animals↗
PubMed1999

Developmental changes in the activation properties and ultrastructure of fast- and slow-twitch muscles from fetal sheep.

At early stages of muscle development, skeletal muscles contract and relax slowly, regardless of whether they are destined to become fast- or slow-twitch. In this study, we have characterised the activation profiles of developing fast- and slow-twitch muscles from a precocial species, the sheep, to determine if the activation profiles of the muscles are characteristically slow when both the fast- and slow-twitch muscles have slow isometric contraction profiles. Single skinned muscle fibres from the fast-twitch flexor digitorum longus (FDL) and slow-twitch soleus muscles from fetal (gestational ages 70, 90, 120 and 140 days; term 147 days) and neonatal (8 weeks old) sheep were used to determine the isometric force-pCa (pCa = -log10[Ca2+]) and force-pSr relations during development. Fast-twitch mammalian muscles generally have a greatly different sensitivity to Ca2+ and Sr2+ whereas slow-twitch muscles have a similar sensitivity to these divalent cations. At all ages studied, the force-pCa and force-pSr relations of the FDL muscle were widely separated. The mean separation of the mid-point of the curves (pCa50-pSr50) was approximately 1.1. This is typical of adult fast-twitch muscle. The force-pCa and force-pSr curves for soleus muscle were also widely separated at 70 and 90 days gestation (pCa50-pSr50 approximately 0.75); between 90 days and 140 days this separation decreased significantly to approximately 0.2. This leads to a paradoxical situation whereby at early stages of muscle development the fast muscles have contraction dynamics of slow muscles but the slow muscles have activation profiles more characteristic of fast muscles. The time course for development of the FDL and soleus is different, based on sarcomere structure with the soleus muscle developing clearly defined sarcomere structure earlier in gestation than the FDL. At 70 days gestation the FDL muscle had no clearly defined sarcomeres. Force (N cm-2) increased almost linearly between 70 and 140 days gestation in both muscle types and there was no difference between the Ca(2+)- and Sr(2+)-activated force throughout development.

Animals↗
PubMed1999

Effects of hyperthermia on uterine blood flow and shunting through uterine arteriovenous anastomoses in the late-pregnant ewe.

The effect of maternal hyperthermia on uterine blood flow (UBF) through the two main uterine arteries and on the proportion of UBF shunted through uterine arteriovenous anastomoses (AVAs) was investigated. Eight late-pregnant ewes were exposed to normothermic (22-23 degrees C) or hyperthermic (approximately 39 degrees C) ambient conditions for 8 h. UBF was measured in the left and right uterine arteries using flow probes and microspheres were injected into the uterine artery before, during and after the experimental period. The distribution of microspheres between the uterus and lungs was determined to calculate changes in capillary and AVA blood flows. Hyperthermia produced a significant (P<0.05) increase in maternal core temperature (+1.5 degrees C), increase in maternal blood pH (+0.21; P<0.05) and decrease in maternal pCO2 (-16.2 mmHg; P<0.05). Blood flow to the uterine horn ipsilateral to the corpus luteum (CL) remained unchanged during hyperthermia, whereas total UBF and blood flow to the contralateral uterine horn were significantly decreased (P<0.05), by 23.1% and 20.8%, respectively, of pre-heat control values. The proportion of UBF shunted through uterine AVAs during hyperthermia was not significantly different from values observed in normothermic ewes (21.9 +/- 0.7%). Mild to moderate hyperthermia in late-pregnant sheep induces respiratory alkalosis and decreases total blood flow to the uterus, brought about by a decrease in blood flow to the uterine horn contralateral, but not ipsilateral to the CL. Heat treatment does not alter the proportion of UBF traversing uterine AVAs.

Animals↗
PubMed1999

Neonatal ethanol exposure alters bcl-2 family mRNA levels in the rat cerebellar vermis.

BACKGROUND: The objective of the present work was to determine whether ethanol-induced cerebellar cell death during development is related to alterations in the expression of bcl-2 family genes. METHODS: Rats were exposed to ethanol or control conditions during the neonatal period and transcript levels of bcl-2 family members relative to cyclophilin were determined. Pups exposed in parallel were taken for cerebellar cell counts. RESULTS: Ethanol exposure during the first postnatal week significantly reduced Purkinje and granule cell numbers by postnatal day 21 (P21). Acute first postnatal week ethanol exposure up-regulated mRNA transcripts encoding the cell death-promoting molecules bax and bcl-xs as measured on P4. An additional day of exposure on P5 resulted in no further alterations in bcl-2 family transcripts, likely because Purkinje cell death was detectable as early as P5. To determine whether proapoptotic gene expression changes were specific to first postnatal week ethanol neurotoxicity, we examined bcl-2 family mRNA levels in rats exposed to ethanol during a developmental period of cerebellar insusceptibility, the second postnatal week. Exposure on P7 to P8 produced no change in cerebellar cell number, but also resulted in increased levels of bax, although only after 2-day ethanol exposure and not after acute exposure on P7. CONCLUSIONS: These data implicate altered expression of proapoptotic members of the bcl-2 gene family in acute ethanol-mediated cerebellar cell death during the first postnatal week. They also suggest that the differential survival of cerebellar neurons after ethanol exposure during more mature developmental stages may be related to more successful suppression of proapoptotic processes.

Animals↗
PubMed1999

BDNF and NGF afford in vitro neuroprotection against ethanol combined with acute ischemia and chronic hypoglycemia.

Consumption of alcohol during pregnancy can result in central nervous system deficits in infants ranging from fetal alcohol effects to fetal alcohol syndrome. Changes in cerebral metabolism causing ischemic in utero conditions can also result from ethanol (EtOH). Growth factors have been shown to ameliorate ischemic damage and EtOH-induced neurotoxicity. However, using an in vitro model system of fetal alcohol effects/fetal alcohol syndrome, this study examines the neuroprotective effects of nerve growth factor, brain-derived neurotrophic factor, or glial cell line derived neurotrophic factor against EtOH treatment (0, 200, 400, 800, or 1, 600 mg/dl) combined with acute ischemia (2-hour hypoxia in EtOH-containing glucose-free media) followed by chronic hypoglycemia (16-hour glucose deprivation in EtOH-containing media). 3-(4, 5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assays assessed relative neurotoxicity. Glial cell derived neurotrophic factor was not neuroprotective. Nerve growth factor protected against ischemia/hypoglycemia combined with 0-1,600 mg/dl EtOH. Brain-derived neurotrophic factor protected against ischemia/hypoglycemia combined with 0-800 mg/dl EtOH. These studies demonstrate marked growth factor neuroprotection against a myriad of conditions encountered by developing EtOH-exposed fetuses.

Acute Disease↗
PubMed1999

Prenatal ethanol exposure reduces spinal cord motoneuron number in the fetal rat but does not affect GDNF target tissue protein.

Fetal rats were exposed throughout gestation to one of three diets: an ethanol-containing liquid diet, a liquid diet with the isocaloric substitution of sucrose for ethanol or a laboratory chow control diet. At postnatal day 1 (P1), the spinal cords were taken for analyses of motoneuron number and size. These analyses revealed a significant loss of motoneurons and a reduction of motoneuron size in the ethanol-exposed animals, compared to both sucrose and chow controls. Spinal cord length and ventral horn volume were not altered as a result of ethanol treatment, so the change in motoneuron number cannot be attributed to volumetric changes. The content of the motoneuron survival factor glial cell-line-derived neurotrophic factor (GDNF) was also assessed in the P1 limb motoneuron target tissue. This analysis was undertaken because GDNF is a potent survival factor for developing motoneurons and has been shown to protect this population from ethanol neurotoxicity. Thus, its depletion could contribute to motoneuron loss. These analyses, using the ELISA assay, did not detect reductions in GDNF in the ethanol-exposed animals. Therefore, alterations in other neurotrophic factors or ethanol neurotoxicity by other means appear to be responsible for the motoneuron loss. These results are consistent with earlier studies in the chick embryo, which also found reduced motoneuron numbers as a function of developmental ethanol exposure, and point again to the general lethality of ethanol to the developing nervous system.

Animal Nutritional Physiological Phenomena↗
PubMed1999

Effects of the respiratory stimulant almitrine on breathing and FOS expression in the brain of fetal and newborn sheep.

Almitrine is a piperazine derivative known to stimulate breathing in the adult but cause apnea in fetal sheep. In fetal sheep (127-133 d gestation; term = 147 d) we confirmed this finding, but found that almitrine (4 mg/kg, i.v. or intra-arterial) had a biphasic effect, briefly stimulating and then suppressing breathing movements for at least 3 h. In 2- to 3-d-old (n = 4) and 7- to 14-d-old (n = 4) lambs almitrine increased both tidal volume and breath frequency, increased arterial partial pressure of oxygen and pH, and decreased partial pressure of carbon dioxide. The changes of tidal volume, partial pressure of oxygen and partial pressure of carbon dioxide were less in the 2- to 3-d-old compared with the 7- to 14-d-old lambs. The distribution of the nuclear phosphoprotein FOS, a marker of neuronal activation was examined in fetal and newborn brains. FOS protein was increased in cardiorespiratory areas of the medulla and pons, in the periaqueductal region of the midbrain, and in the supraoptic and paraventricular regions of the hypothalamus. In the pons, FOS protein was increased in the medial parabrachial and subcoeruleus nuclei in the fetuses but not in the 2- to 3- or 7- to 14-d-old lambs. These observations are similar to those reported for hypoxia, and consistent with the hypothesis that both almitrine and hypoxia inhibit fetal breathing movements by an action on a select group of pontine neurons. Whether these neurons respond directly to these stimuli or receive input from the other centers is yet to be elucidated. The mechanisms that change the almitrine (and hypoxia) response from inhibition to excitation at birth have not been identified, but may be important in preventing apnea in the newborn.

Almitrine↗
PubMed1999

Kynurenic acid in brain and cerebrospinal fluid of fetal, newborn, and adult sheep and effects of placental embolization.

Concentrations of the endogenous glutamate receptor antagonist kynurenic acid (KA) were measured in various brain regions and in cisternal cerebrospinal fluid of fetal, newborn, and adult sheep. KA concentrations were significantly higher in the fetal brain and cerebrospinal fluid at 90 and 140 d gestation compared with postnatal ages. In fetuses of 132-139 d gestation, KA concentrations in cerebrospinal fluid collected by drainage from an indwelling cisternal catheter increased significantly after infusion of the organic acid transport inhibitor probenecid (100 or 200 mg/kg, i.v.) indicating active transport of KA out of the fetal brain. In fetuses in which the umbilical circulation had been chronically restricted from 120 to 140 d gestation by partial embolization of the placenta, plasma concentrations of the KA precursor kynurenine were significantly lower than in control fetuses, and KA concentrations in the hypothalamus and hippocampus were significantly reduced; other brain regions were not affected. These results indicate that the production of KA is higher in the fetal brain compared with the newborn and adult brain. Because KA diminishes the risk of excitotoxic neuronal damage under hypoxic-ischemic conditions, the high levels of KA in the brain before birth may have a neuroprotective function. The decrease of KA concentrations in the hypothalamus and hippocampus after umbilical embolization suggests that, after chronic hypoxia in utero, these regions of the brain may become more vulnerable to subsequent episodes of acute hypoxia or ischemia encountered in late gestation or during parturition.

Animals↗
PubMed1999

Chronic ethanol exposure increases 3H-GABA release in rat hippocampus by presynaptic muscarinic receptor modulation.

BACKGROUND: Chronic ethanol treatment (CET) for 28 weeks significantly increases electrically-stimulated 3H-GABA release from hippocampal slices. This increase in GABA release may be one of the mechanisms by which CET decreases the magnitude of long-term potentiation (LTP) in the hippocampus. The present study examined whether CET increases GABA release via an alteration in heterologous presynaptic cholinergic regulation. METHODS: Animals were treated with ethanol or sucrose diet for 28 weeks followed by either no withdrawal or a 48-hr withdrawal period. The electrically-stimulated 3H-GABA release from preloaded superfused hippocampal slices of naive and CET rats was measured. RESULTS: Carbachol increased 3H-GABA release in a concentration-dependent manner, and atropine modulated 3H-GABA release in a biphasic concentration-dependent manner. Atropine (10 microM) significantly blocked the effects of carbachol. Oxotremorine, a selective muscarinic receptor agonist, also increased 3H-GABA release. Mecamylamine, a selective nicotinic antagonist, did not modulate 3H-GABA release and did not block the effects of carbachol. The effects of these agents were also tested in rats 0 or 48 hrs after withdrawal from CET. The biphasic effects of atropine were decreased, whereas the facilitating effects of carbachol were significantly increased. There were no changes in the effects of these agents on 3H-acetylcholine release from hippocampal slices of CET rats compared to sucrose-treated rats. CONCLUSION: These results suggest that presynaptic muscarinic receptors facilitate GABA release, whereas nicotinic receptors do not play a significant role in modulating GABA release in hippocampus. CET selectively alters presynaptic muscarinic regulation of GABA release in hippocampus and may help us to further understand the mechanism underlying the disruption of LTP by CET.

Animals↗
PubMed1998

Effect of pregnane steroids on electrocortical activity and somatosensory evoked potentials in fetal sheep.

The effect of infusing the neuroactive steroids pregnanolone and iso-pregnanolone on somatosensory evoked potentials (SEP) and electrocortical (ECoG) activity was studied in unanaesthetised fetal sheep, 130-135 days gestation. Intravenous infusion of pregnanolone (6 mg/kg per h) significantly increased the proportion of high voltage ECoG (56.1+/-4.8% vs. control 43.5+/-3.2%, P < 0.05), and decreased low voltage ECoG (43.9+/-4.8% vs. control 56.6+/-3.2%, P < 0.05). Pregnanolone treatment decreased the amplitude of the N25 peak of the SEP (89.9+/-2.8% of control, P < 0.05) evoked following stimulation of the skin of the upper lip. In contrast, iso-pregnanolone treatment had no effect on ECoG activities, or on the amplitude and latency of peaks in the SEP. We conclude that 3alpha-hydroxy pregnane steroids are active at GABA(A) receptors in fetal sheep and can modulate sleep/wake activity before birth.

Animals↗
PubMed1998

Prenatal ethanol exposure reduces parvalbumin-immunoreactive GABAergic neuronal number in the adult rat cingulate cortex.

The effect of prenatal ethanol exposure on the number of parvalbumin-immunoreactive (PA+) GABAergic neurons in the adult rat anterior cingulate cortex was determined. Pregnant Long-Evans rats were maintained on one of three diets throughout gestation: an ethanol-containing liquid diet, a similar, control liquid diet with the isocaloric substitution of sucrose for ethanol, or a lab chow control diet. Offspring were euthanized on postnatal day 60 and brains were prepared for parvalbumin immunocytochemistry. Rats exposed to the ethanol-containing diet contained 45% fewer PA+ neurons in the anterior cingulate cortex compared with sucrose and chow controls. This reduction occurred in the absence of changes in structure volume, and occurred in the absence of changes in PA+ neuronal size.

Animals↗
PubMed1998

A comparative study of ethanol, hypoglycemia, hypoxia and neurotrophic factor interactions with fetal rat hippocampal neurons: a multi-factor in vitro model developmental ethanol effects.

Fetal alcohol syndrome (FAS) is characterized by numerous central nervous system anomalies, with the hippocampus being particularly vulnerable to developmental ethanol exposure. In addition to direct ethanol neurotoxicity, other conditions resulting from maternal ethanol consumption, such as hypoglycemia and hypoxia, may also contribute to FAS. The present study used a tissue culture system to model multiple conditions which may relate to in vivo FAS, and assessed their relative neurotoxicity with MTT assays. Gestational day 18 rat hippocampal cultures were exposed to varying ethanol concentrations, glucose withdrawal-induced hypoglycemic (gwHG, 16 h) or acute hypoxic (aHP, 2 h) conditions alone, as well as to co-treatments with ethanol and gwHG or aHP. Brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) have previously been shown to ameliorate ethanol-, hypoglycemia- and hypoxia-induced neurotoxicity. Therefore, their neuroprotective potential, along with ciliary neurotrophic factor (CNTF), was examined. Neuronal viability was reduced dose-dependently by ethanol, alone or with hypoglycemia or hypoxia. Ethanol + gwHG or aHP was not uniformly additive. NGF treatment provided the most extensive neuroprotection, being effective against ethanol (200 and 400 mg/dl), gwHG, and aHP, alone and combined. BDNF afforded similar protection, but not against ethanol + gwHG. CNTF protected only against aHP. CNTF + BDNF, previously shown to act synergistically, protected against ethanol + aHP up to 800 mg/dl ethanol, but not, paradoxically, against ethanol alone, gwHG, or ethanol + gwHG, all conditions BDNF alone protected against. This study demonstrated that several neurotrophic factors are capable of mitigating neurotoxicity associated with ethanol, hypoglycemia and hypoxia.

Animals↗
PubMed1998

Effect of probenecid on breathing movements and cerebral clearance of prostaglandin E2 in fetal sheep.

1. Intravenous infusion of probenecid (79-160 mg kg-1) into unanaesthetized fetal sheep (127-143 days gestation) in utero significantly decreased the incidence and amplitude of spontaneous breathing movements, but did not change the incidence of low voltage electrocortical (ECoG) activity, plasma prostaglandin E2 (PGE2) concentrations, blood gases or pH. 2. In fetuses pretreated with paracetamol (350 mg kg-1) to inhibit PG synthase activity, infusion of probenecid did not change the mean incidence or amplitude of breathing movements, indicating that the inhibitory effect of probenecid on breathing movements required the presence of active PG synthesis. 3. Probenecid infusion in four unanaesthetized fetuses significantly increased the PGE2 concentrations in cisternal cerebrospinal fluid (CSF) by 6.6 +/- 1.5-fold (P < 0.05). 4. In pentobarbitone-anaesthetized, exteriorized fetuses, probenecid infusion decreased the clearance of [3H]PGE2 from CSF during ventriculo-cisternal perfusion of artificial CSF containing [3H]PGE2. 5. These results suggest that there is active transport of PGs from CSF to blood in fetal sheep from at least 127 days gestation. Inhibition of this transport results in the accumulation of PGs within interstitial fluid of the brain, one effect of which is to suppress the spontaneous activity of the respiratory centres.

Anesthesia, General↗